Mechanism of membrane fusion by fusion protein

融合蛋白的膜融合机制

• 批准号: 11694096
• 负责人: NAITO Akira
• 金额: $ 2.43万
• 依托单位: Himeji Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11694096/
• 关键词: Influenza virus; Solid state NMR; Fusion peptide; lipid bilayer; α-helix; Neutron scattering; Chemical shifts; Biomembrane; 膜融合活性蛋白質; プロトンポンプ; メリチン; αーヘリックス; ^<31>P NMR; 磁場配向

In this research project, we have studied to understand the molecular mechanism of viral infectn using influenza fusion peptides. We have particularly focused to determine the structure and orientation of the peptides bund to the membrane. Following results were obtained in this research project.(1) The conformation and dynamics of melittin bound to the dimyristoylphophatidylcholin (DMPC) bilayer and the magnetic orientation in the lipid bilayer systems were investigated by solid-state ^<31>P and ^<13>C NMR spectroscopy. Using ^<31>P NMR, it was found that melittin-DMPC bilayer system forms magnetically oriented elongated vesicles with the long axis parallel to the magnetic field above the liquid crystalline-gel phase transition temperature. ^<13>C NMR spectra were observed on the ^<13>C labeled melittin bound to the lipid bilayers. Finally, it was found that melittin adopts a transmembrane α-helix whose average axis is parallel to the bilayer normal.(2) N-terminal fragment of HA2 (1-27) in influenza virus was synthesized and investigated by solid state NMR spectroscopy. It was found that the N-terminal part of the fragment forms α-helix and showed different behavior of interaction with membrane at acidic pH from that at basic pH condition.(3) Transmembrane fragment of M2 protein of influenza virus (M2-TMP) was synthesized and investigated by means of ^<15>N solid state NMR spectroscopy. The results indicate that the α-helical axis is tilting by 33° and 37° from the bilayer normalin the DOPC and DMPC bilayer systems, respectively.(4) Membran insertion behavior of influenza fusion peptide was investigated by a neutron scattering method. The results indicate that the helical axis is tilting by 55° from the bilayer normal and the helix is located around the surface of bilayers.

在该研究项目中，我们研究了使用影响天堂来了解病毒感染的分子机制。我们特别专注于确定肽束的结构和方向。 （1）通过固体 ^<31> p和 ^<31> p和 ^<13> c nmr光谱法研究了该研究项目中的结果。使用 ^<31> p NMR，发现Melittin-DMPC双层系统形成磁取向的细长蔬菜，长轴平行于液体结晶凝胶相变温度上方的磁场。 ^<13> C NMR光谱在与脂质双层结合的蜂胶蛋白标记的 ^<13> C上观察到。最后，发现蜂毒素采用了跨膜α-螺旋，其平均轴与双层的平均轴平行。（2）HA2（1-27）流感病毒的N末端片段已通过固体状态NMR光谱合成并研究。发现片段形式的N末端部分α-螺旋在基本pH条件下显示出与酸性pH的膜相互作用不同的行为。结果表明，α-螺旋轴分别从双层轴倾斜33°和37°，分别是DOPC和DMPC双层系统。（4）通过中子散射方法研究了影响Za融合肽的膜插入行为。结果表明，螺旋轴与双层正常的倾斜55°，并且螺旋位于双层表面周围。

项目成果

K.Nishimura: "Natural abundance ^<13>C REDOR coupled to a singly ^<15>N-labeled nucleus : simulaneous determination of interatomic distances in crystalline ammonium [^<15>N] L-glutamate monohydrate"J.Mol.Struct.. 560. 29-38 (2001)

K.Nishimura：“天然丰度^ 13 C REDOR与单个^ 15 N标记核偶联：同时测定结晶铵[^ 15 N] L-谷氨酸一水合物中的原子间距离”J.Mol。

H.Saito: "Conformation and backbone dynamics of bacteriorhodopsin revealed by ^<13>C NMR"Biochim.Biophys.Acta. 1460. 39-48 (2000)

H.Saito：“通过 13 C NMR揭示的细菌视紫红质的构象和主链动力学”Biochim.Biophys.Acta。

S.Tuzi: "Localization of a Cation Binding Site in the Loop between Helices F and G of Bacteriorhodopsin, as Studied by ^<13>C NMR,"Biophys.J.. 76. 1523-1531 (1999)

S.Tuzi：“细菌视紫红质的螺旋F和G之间的环中的阳离子结合位点的定位，通过13C NMR研究”，Biophys.J.. 76. 1523-1531(1999)

M.Kamihira: "Phenyl ring dynamics of enkephalin molecules and behavior of bound solvents in the crystalline states by ^2H NMR spectroscopy"J.Phys.Chem.. 103. 3356-3363 (1999)

M.Kamihira：“脑啡肽分子的苯环动力学和结晶状态下结合溶剂的行为，通过^2H NMR 光谱”J.Phys.Chem.. 103. 3356-3363 (1999)

J.Kimura: "Determination of N-H bond lengths of ^<15>N-labeled poly (L-alanines) by ^1H CRAMPS NMR"Macromolecules. 18. 6627-6629 (2000)

J.Kimura：“通过1H CRAMPS NMR测定15 N-标记的聚(L-丙氨酸)的N-H键长度”大分子。